Burke T W, Kadonaga J T
Department of Biology and Center for Molecular Genetics, University of California, San Diego, La Jolla, California 92093-0347 USA.
Genes Dev. 1997 Nov 15;11(22):3020-31. doi: 10.1101/gad.11.22.3020.
We analyzed the function of the downstream promoter element (DPE), a distinct 7-nucleotide core promoter element that is approximately 30 nucleotides downstream of the transcription start site of many TATA-box-deficient (TATA-less) promoters in Drosophila. There is a strict requirement for spacing between the Inr and DPE motifs, as an increase or decrease of 3 nucleotides in the distance between the Inr and DPE causes a seven- to eightfold reduction in transcription as well as a significant reduction in the binding of purified TFIID. These results suggest a specific and somewhat rigid interaction of TFIID with the Inr and DPE sequences. Photo-cross-linking analysis of purified TFIID with a TATA-less DPE-containing promoter revealed specific cross-linking of dTAFII60 and dTAFII40 to the DPE, with a higher efficiency of cross-linking to dTAFII60 than to dTAFII40. These data, combined with the previously well-characterized interactions between the two TAFs and their homology to histones H4 and H3, suggest that a dTAFII60-dTAFII40 heterotetramer binds to the DPE. Human and Drosophila transcription factors exhibit essentially the same requirements for DPE sequence and for Inr-DPE spacing. In addition, the TATA-less promoter of the human interferon regulatory factor-1 (IRF-1) gene contains a DPE that is important for transcriptional activity both in vitro and in cultured cells. Hence, these studies provide evidence for a direct role of TAFs in basal transcription of TATA-less DPE-containing genes and collectively indicate that the DPE is, in many respects, a downstream counterpart to the TATA box that is present in Drosophila to humans.
我们分析了下游启动子元件(DPE)的功能,DPE是一种独特的7核苷酸核心启动子元件,位于果蝇许多缺乏TATA盒(无TATA)启动子转录起始位点下游约30个核苷酸处。Inr和DPE基序之间的间距有严格要求,因为Inr和DPE之间的距离增加或减少3个核苷酸会导致转录降低7至8倍,以及纯化的TFIID结合显著减少。这些结果表明TFIID与Inr和DPE序列存在特定且略显刚性的相互作用。对纯化的TFIID与含无TATA DPE的启动子进行光交联分析,揭示了dTAFII60和dTAFII40与DPE的特异性交联,与dTAFII40相比,与dTAFII60的交联效率更高。这些数据,结合之前对这两种TAF之间充分表征的相互作用及其与组蛋白H4和H3的同源性,表明dTAFII60 - dTAFII40异源四聚体与DPE结合。人类和果蝇转录因子对DPE序列和Inr - DPE间距表现出基本相同的要求。此外,人类干扰素调节因子1(IRF - 1)基因的无TATA启动子含有一个DPE,其对体外和培养细胞中的转录活性都很重要。因此,这些研究为TAF在含无TATA DPE基因的基础转录中的直接作用提供了证据,并共同表明DPE在许多方面是果蝇到人类中存在的TATA盒的下游对应物。